This invention relates to a series of 3,3-disubstituted-oxindole derivatives that are useful in the treatment of hyperproliferative diseases, such as cancers, in mammals. This invention also relates to a method of using such compounds in the treatment of hyperproliferative diseases in mammals, especially humans, to pharmaceutical compositions containing such compounds, and to methods of preparing such compounds. Oxindole derivatives alleged to have CNS activity have been described in EP 0311010 B1 and EP 241 006.
Oncogenes frequently encode protein components of signal transduction pathways which lead to stimulation of cell growth and mitogenesis. Oncogene expression in cultured cells leads to cellular transformation, characterized by the ability of cells to grow in soft agar and the growth of cells as dense foci lacking the contact inhibition exhibited by non-transformed cells. Mutation and/or overexpression of certain oncogenes is frequently associated with human cancer.
To become functional, the precursor of the Ras oncoprotein must undergo farnesylation of the cysteine residue located in a carboxyl-terminal tetrapeptide. Inhibitors of the enzyme that catalyzes this modification, farnesyl protein transferase, have therefore been suggested as agents to combat tumors in which Ras contributes to transformation. Mutated, oncogenic forms of Ras are frequently found in many human cancers, most notably in more than 50% of colon and 90% pancreatic carcinomas (Kohl et al., Science, Vol. 260, 1834 to 1837, 1993). The compounds of the present invention exhibit activity as inhibitors of the enzyme farnesyl protein transferase and are therefore believed to be useful as anti-cancer and anti-tumor agents. Further, the compounds of the present invention may be active against any tumors that proliferate by virtue of farnesyl protein transferase.
The present invention relates to compounds of formula 1 
and to pharmaceutically acceptable salts, prodrugs, and solvates thereof wherein:
n is 0 or 1;
R1 is C1-C3 alkylpyridyl or C1-C3 alkylimidazolyl;
R2 is selected from the group consisting of C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, xe2x80x94(CH2)p(C6-C10 aryl), and xe2x80x94(CH2)p(4-10 membered unsaturated heterocyclyl), wherein p is an integer from 0 through 3, and wherein any of said R1 and R2 groups are optionally substituted with 1 to 3 R6 groups;
R3 is xe2x80x94(CH2)m(1- or 2-adamantyl), C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, xe2x80x94(CH2)m(C6-C10 aryl), 
X1, X2, and X3 are each independently C1-C7 alkylene optionally containing 1 or 2 carbon-carbon double bonds, X4 is a bond or C1-7 alkylene optionally containing 1 or 2 carbon-carbon double or triple bonds, and, in formula (1b), the X4 moiety is attached to the X1 moiety at any available carbon in the X1 moiety,
and each of the foregoing R3 groups are optionally substituted with an R5 group and optionally with 1 to 4 R6 groups,
or R3 is xe2x80x94(CH2)tSO2R9, xe2x80x94(CH2)tC(O)R9, or xe2x80x94(CH2)m(4-10 membered heterocyclyl) optionally substituted with 1 to 5 R6 groups;
m, in the aforementioned R3 groups, is independently an integer from 0 through 6 and t is independently an integer from 1 through 5;
R4 is C6-C10 aryl or 4-10 membered heterocyclyl, each of said R4 groups being optionally substituted by 1 to 3 R6 groups;
each R5 is independently selected from halo, C1-C6 alkyl optionally substituted by 1 to 3 halo, nitro, cyano, xe2x80x94OR9, xe2x80x94C(O)R9, SR9, xe2x80x94SO2R9, xe2x80x94SO3H, xe2x80x94S(O)R9, xe2x80x94NR7R8, xe2x80x94CHxe2x95x90NOR7, xe2x80x94C(O)OR9, xe2x80x94OC(O)R9, xe2x80x94SO2NR9R8, xe2x80x94C(O)NR9R8, xe2x80x94NR8C(O)R9, xe2x80x94OC(O)NR9R8, xe2x80x94C(O)ONR7R9, xe2x80x94NR8C(O)NR9R8, xe2x80x94NR8C(O)O(C1-C4 alkyl), xe2x80x94C(NR8)NR9R8, xe2x80x94C(NCN)NR9R8, xe2x80x94C(NCN)S(C1-C4 alkyl or C1-C4 haloalkyl), xe2x80x94NR8C(NCN)S(C1-C4 alkyl or C1-C4 haloalkyl), xe2x80x94NR8C(NCN)NR7R8, xe2x80x94NR8SO2(C1-C4 alkyl or C1-C4 haloalkyl), xe2x80x94NR8C(O)C(O)R8, xe2x80x94NR8C(O)C(O)NR9R8, xe2x80x94P(O)(OR7)2, and xe2x80x94CH2)q(4-10 membered heterocyclyl), each q is independently an integer from 0 through 3, and the alkyl and heterocyclyl moieties of the foregoing R5 groups are optionally substituted by 1 to 3 R10 groups;
each R6 is independently selected from R5, C2-C10 alkenyl, C2-C10 alkynyl and xe2x80x94(CH2)t(C6-C10 aryl) optionally substituted with 1 to 3 R10 groups, t is an integer from 0 through 3;
each R7 is independently hydrogen or C1-C4 alkyl optionally substituted by 1 to 3 halo;
each R8 is independently R7 or xe2x80x94OR7;
each R9 is independently selected from hydrogen, C1-C6 alkyl, xe2x80x94(CH2)q(C6-C10 aryl) and xe2x80x94(CH2)q(4-10 membered heterocyclyl), said R9 groups, except H, are optionally substituted with 1 to 3 R10 groups, and each q is independently an integer from 0 through 3; and,
each R10 is independently selected from halo, nitro, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, xe2x80x94C(O)O(C1-C6 alkyl), and C6-C10 aryl.
Preferably, in the compounds of formula 1, each p integer in R2 is an integer independently selected from 0 to 3, more preferably an integer independently selected from 1 to 3, 1 being most preferred.
Preferred compounds of formula 1 include those wherein one or both of R1 and R2 is xe2x80x94(CH2)p(4-10 membered unsaturated heterocyclyl) optionally substituted with 1 to 3 R6 groups, more preferably xe2x80x94(CH2)p(5 or 6 membered unsaturated heterocyclyl). Preferably, each heterocyclyl of R1 and R2 is independently imidazolyl or pyridinyl. In different embodiments, one or both R1 and R2 is 2-, 3-, or 4-pyridinylmethyl; preferably, one or both of R1 and R2 is 4-pyridinylmethyl. In other embodiments, R1 and R2 are each independently imidazol-1-ylmethyl, imidazol-2-ylmethyl, or imidazol-4-ylmethyl, optionally substituted with 1 to 3 R6 groups; preferably, R1 and R2 are both imidazol-4-ylmethyl, each optionally substituted with 1 to 3 R6 groups. When only one of R1 and R2 is a xe2x80x94(CH2)p(4-10 membered unsaturated heterocyclyl) optionally substituted with 1 to 3 R6 groups, the other of R1 or R2 is a C1-C10 alkyl substituted by one R6 group, wherein the Re group is preferably, xe2x80x94SR9; preferably, both of R1 or R2 is imidazolyl or pyridinyl, more preferably imidazol-1-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl or 4-pyridinylmethyl.
Preferred compounds of formula 1 include those wherein R3 is xe2x80x94(CH2)m(1- or 2-adamantyl) or xe2x80x94(CH2)m(C6-C10 aryl), wherein the aryl group is optionally substituted with 1 to 5 R6 groups, preferably wherein m is an integer 1. Preferably the aryl group is phenyl or naphthyl and R6 is R5, wherein R5 is xe2x80x94SO2R9, xe2x80x94SO2NR9R8, or xe2x80x94C(O)OR9, preferably, xe2x80x94SO2NR9R8.
Other preferred compounds of formula 1 include those wherein R4 is C6-C10 aryl substituted by R6, preferably, wherein the R6 is cyano. Other preferred compounds of formula 1 include those wherein R4 is C6-C10 aryl substituted by R6, wherein the R6 is preferably halo or formyl, provided that when the R6 is bromo, then R3 is substituted by R5, wherein R5 is sulfonamide.
Specific preferred compounds include the following:
4-[6-(4-Cyano-phenyl)-3,3-bis-(1H-imidazol-4-ylmethyl)-2-oxo-2,3-dihydro-indol-1-ylmethyl]-N,N-dimethyl-benzenesulfonamide;
4-[3,3-Bis-(3H-imidazol-4-ylmethyl)-1-naphthalen-1-ylmethyl-2-oxo-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-[1-Adamantan-1-ylmethyl-3-(1H-imidazol-4-ylmethyl)-3-(3H-imidazol-4-ylmethyl)-2-oxo-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-[3-(1H-imidazol-4-ylmethyl)-3-(3H-imidazol-4-ylmethyl)-2-oxo-1-quinolin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-[6-(4-Formyl-phenyl)-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-indol-1-ylmethyl]-N,N-dimethyl-benzenesulfonamide;
4-[6-(4-Cyano-phenyl)-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-indol-1-ylmethyl]-N,N-dimethyl-benzenesulfonamide;
4-(1-Naphthalen-1-ylmethyl-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl)-benzonitrile;
4-(2-Oxo-3,3-bis-pyridin-4-ylmethyl-1-quinolin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl)-benzonitrile;
4-[1-Adamantan-1-ylmethyl-3,3-bis-(3-methyl-3H-imidazol-4-ylmethyl)-2-oxo-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-(7-Methyl-1-naphthalen-2-ylmethyl-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl)-benzonitrile;
4-(7-Methyl-1-naphthalen-1-ylmethyl-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl)-benzonitrile;
4-[6-(4-Cyano-phenyl)-7-methyl-2-oxo-3,3-bis-pyridin-4-ylmethyl-2,3-dihydro-indol-1-ylmethyl]-N,N-dimethyl-benzenesulfonamide;
4-[7-Methyl-1-naphthalen-1-ylmethyl-2-oxo-3-(1H-pyrazol-4-ylmethyl)-3-pyridin-4-ylmethyl-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-[3,3-Bis-(1H-imidazol-4-ylmethyl)-7-methyl-1-naphthalen-1-ylmethyl-2-oxo-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-[7-Methyl-3,3-bis-(5-methyl-1H-imidazol-4-ylmethyl)-1-naphthalen-1-ylmethyl-2-oxo-2,3-dihydro-1H-indol-6-yl]-benzonitrile;
4-(2-Naphthalen-1-ylmethyl-1,3-dioxo-4,4-bis-pyridin-4-ylmethyl-1,2,3,4-tetrahydro-isoquinolin-7-yl)-benzonitrile; and
4-{1,3-Dioxo-4,4-bis-pyridin-4-ylmethyl-2-[1-(thiophene-2-sulfonyl)-pyrrolidin-3-yl]-1,2,3,4-tetrahydro-isoquinolin-7-yl}-benzonitrile.
and the pharmaceutically acceptable salts, prodrugs, and solvates of the foregoing compounds, as well as stereoisomers of the foregoing compounds.
This invention also relates to a method for the treatment of abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of a compound of the formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug or solvate thereof, that is effective in inhibiting farnesyl protein transferase. In one embodiment of this method, the abnormal cell growth is cancer, including, but not limited to, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin""s Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, or a combination of one or more of the foregoing cancers. In another embodiment of said method, said abnormal cell growth is a benign proliferative disease, including, but not limited to, psoriasis, benign prostatic hypertrophy or restinosis.
This invention also relates to a method for the treatment of abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of a compound of the formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, that is effective in treating abnormal cell growth.
This invention also relates to a method for the treatment of abnormal cell growth in a mammal which comprises administering to said mammal a therapeutically effective amount of a compound of formula 1, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, in combination with an anti-tumor agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.
The present invention also relates to a method for the treatment of an infection in a mammal, including a human, that is facilitated by farnesyl protein transferase, such as hepatitis delta virus or malaria, which comprises administering to said mammal a therapeutically effective amount of a compound of formula 1 or a pharmaceutically acceptable salt, prodrug, or solvate thereof.
This invention also relates to a pharmaceutical composition for the treatment of abnormal cell growth in a mammal, including a human, comprising an amount of a compound of the formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, that is effective in inhibiting farnesyl protein transferase, and a pharmaceutically acceptable carrier. In one embodiment of said composition, said abnormal cell growth is cancer, including, but not limited to, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin""s Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft issue, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, or a combination of one or more of the foregoing cancers. In another embodiment of said pharmaceutical composition, said abnormal cell growth is a benign proliferative disease, including, but not limited to, psoriasis, benign prostatic hypertrophy or restinosis.
This invention also relates to a pharmaceutical composition for the treatment of abnormal cell growth in a mammal, including a human, comprising an amount of a compound of the formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, that is effective in treating abnormal cell growth, and a pharmaceutically acceptable carrier.
The invention also relates to a pharmaceutical composition for the treatment of abnormal cell growth in a mammal, including a human, which comprises a therapeutically effective amount of a compound of formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, in combination with a pharmaceutically acceptable carrier and an anti-tumor agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.
This invention also relates to a pharmaceutical composition for the treatment of an infection in a mammal, including a human, that is facilitated by farnesyl protein transferase, such as malaria or hepatitis delta virus, comprising an amount of a compound of the formula 1, as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, that is effective in treating abnormal cell growth, and a pharmaceutically acceptable carrier.
xe2x80x9cAbnormal cell growthxe2x80x9d, as used herein, unless otherwise indicated, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the abnormal growth of (1) tumor cells (tumors) expressing an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated as a result of oncogenic mutation in another gene; (3) benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs; and (4) any tumors that proliferate by virtue of farnesyl protein transferase.
The term xe2x80x9ctreatingxe2x80x9d, as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term xe2x80x9ctreatmentxe2x80x9d, as used herein, unless otherwise indicated, refers to the act of treating as xe2x80x9ctreatingxe2x80x9d is defined immediately above.
The term xe2x80x9chaloxe2x80x9d, as used herein, unless otherwise indicated, means fluoro, chloro, bromo or iodo. Preferred halo groups are fluoro, chloro and bromo.
The term xe2x80x9calkylxe2x80x9d, as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight or branched moieties.
The term xe2x80x9ccycloalkylxe2x80x9d, as used herein, unless otherwise indicated, includes cyclic alkyl moieties wherein alkyl is as defined above.
The term xe2x80x9calkenylxe2x80x9d, as used herein, unless otherwise indicated, includes alkyl moieties having at least one carbon-carbon double bond wherein alkyl is as defined above.
The term xe2x80x9calkynylxe2x80x9d, as used herein, unless otherwise indicated, includes alkyl moieties having at least one carbon-carbon triple bond wherein alkyl is as defined above.
The term xe2x80x9calkylenexe2x80x9d, as used herein, unless otherwise indicated, means divalent hydrocarbon radicals which are straight or branched.
The term xe2x80x9calkoxyxe2x80x9d, as used herein, unless otherwise indicated, includes O-alkyl groups wherein alkyl is as defined above.
The term xe2x80x9carylxe2x80x9d, as used herein, unless otherwise indicated, includes an organic radical derived from an aromatic hydrocarbon by removal of one hydrogen, such as phenyl or naphthyl.
The term xe2x80x9c4-10 membered heterocyclicxe2x80x9d, as used herein, unless otherwise indicated, includes aromatic and non-aromatic heterocyclic groups containing one or more heteroatoms, generally 1 to 4 heteroatoms, each selected from O, S and N, wherein each heterocyclic group has from 4-10 atoms in its ring system. Non-aromatic heterocyclic groups include groups having only 4 atoms in their ring system, but aromatic heterocyclic groups must have at least 5 atoms in their ring system. The heterocyclic groups include benzo-fused ring systems and ring systems substituted with one or more oxo moieties. An example of a 4 membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5 membered heterocyclic group is thiazolyl and an example of a 10 membered heterocyclic group is quinolinyl. Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. The foregoing groups, as derived from the compounds listed above, may be C-attached or N-attached where such is possible. For instance, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).
The term xe2x80x9cpharmaceutically acceptable salt(s)xe2x80x9d, as used herein, unless otherwise indicated, includes salts of acidic or basic groups that may be present in the compounds of formula 1. The compounds of the present invention that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that may be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, L!L., salts containing pharmacologically acceptable anions, such as the hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, hydrogen phosphate, dihydrogen phosphate, isonicotinate, acetate, lactate, sailcylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamaite, methanesulfonate (meslyate), ethanesulfonate, benzenesulfanate, p-toluenesulfonate (tosylate), mandelate, and pamoate i.e., 1,1xe2x80x2-methylene-bis-(2-hydroxy-3-naphthoate) salts. The compounds of the present invention that include a basic moiety, such as an amino group, may form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above.
Those compounds of the present invention that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include the alkali metal or alkaline earth metal salts and, particularly, the calcium, magnesium, sodium and potassium salts of the compounds of the present invention.
Certain compounds of formula 1 may have asymmetric centers and therefore exist in different enantiomeric and diastereomeric forms. All optical isomers and stereoisomers of the compounds of formula 1, and mixtures thereof, and all pharmaceutical compositions and methods of treatment that may employ or contain them, are considered to be within the scope of the invention. With respect to the compounds of formula 1, the invention includes the use of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, or mixtures thereof. In particular, the carbon to which the R1 and R2 groups are attached represents a potential chiral center; the present invention encompasses all stereoisomers based on this chiral center. The compounds of formula 1 may also exist as tautomers. This invention relates to the use of all such tautomers and mixtures thereof. Certain compounds of formula 1 may also include oxime moieties, such as where R5 is xe2x80x94CHxe2x95x90NOR7, that exist in E or Z configurations. The present invention includes racemic mixtures of compounds of formula 1 that include such oxime moieties or specific E or Z isomers of such compounds.
This invention also includes isotopically-labelled compounds, and the pharmaceutically acceptable salts thereof, which are identical to those recited in formula 1, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 18O 17O, 35S, 18F, and 36Cl, respectively. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as 3H and 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labelled compounds of formula 1 of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples and Preparations below, by substituting a readily available isotopically labelled reagent for a non-isotopically labelled reagent.
This invention also encompasses pharmaceutical compositions containing and methods of treating hyperproliferative diseases through administering prodrugs of compounds of the formula 1. Compounds of formula 1 having free amino, amido, hydroxy, or carboxylic groups can be converted into prodrugs. Prodrugs include compounds wherein an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues is covalently joined through an amide or ester bond to a free amino, hydroxy or carboxylic acid group of compounds of formula 1. The amino acid residues include but are not limited to the 20 naturally occurring amino acids commonly designated by three letter symbols and also includes 4-hydroxyproline, hydroxylysine, demosine, isodemosime, 3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid, citrulline homocysteine, homoserine, ornithine and methionine sulfone.
Additional types of prodrugs, are also encompassed. For instance, free carboxyl groups can be derivatized as amides or alkyl esters. The amide and ester moieties may incorporate groups including but not limited to ether, amine and carboxylic acid functionalities. Free hydroxy groups may be derivatized using groups including but not limited to hemisuccinates, phosphate esters, dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlined in D. Fleisher, R. Bang, B. H. Stewart, Advanced Drug Delivery Reviews 19, p. 115 (1996). Carbamate prodrugs of hydroxy and amino groups are also included, as are carbonate prodrugs and sulfate esters of hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may be an alkyl ester, optionally substituted with groups including but not limited to ether, amine and carboxylic acid functionalities, or where the acyl group is an amino acid ester as described above, are also encompassed. Prodrugs of this type are described in R. P. Robinson et al., J. Med. Chem. 39, p. 10 (1998). Introduction of prodrug side chains that can be carried out on the hydroxy groups of the compounds of formula 1. For instance, silylation, acylation, alkylation, etc. can be carried out on hydroxy groups of the compounds of formula 1. Selective introduction of prodrug side chains can be carried out on the compounds of formula 1 having more that one protectible group. For example, derivatization of one hydroxy group of a polyhydoxylated compound of formula 1 may be carried out.
The compounds of formula 1 may be prepared as described below.
With reference to Scheme 1 below, the compounds of formula 1, wherein the R1 and R2 substituents are the same and are designated by R, may be prepared by alkylating an intermediate oxindole of formula 2, according to methods familiar to those skilled in the art, such as by stirring the intermediate of formula 2 in the presence of a base and an appropriate alkylating agent, such as Rxe2x80x94Z, wherein Z is a leaving group, such as chloro in an appropriate solvent. An appropriate base is, for example, potassium hexamethyldisilazide. An appropriate solvent is, for example, tetrahydofuran. 
With reference to Scheme 2 below, the compounds of formula 1, may be prepared by coupling a compound of formula 3, wherein W is an appropriate leaving group, such as halo with a compound of the formula ZR4, wherein R4 is as defined above and Z is an appropriate leaving group, such as xe2x80x94B(OH)2. Said reaction requires the presence of a coupling reagent. Coupling reagents familiar to those skilled in the art include metallic and organometallic reagents. An example of an organometallic reagent is tetrakis(triphenylphosphine)palladium (0), which may be used in the presence of an appropriate solvent, such as toluene/ethanol, and the presence of an aqueous base at a temperature ranging from room temperature to 120xc2x0 C., preferably from about 80 to 100xc2x0 C. 
With reference to Scheme 3 below, compounds of formula 1 can be prepared by reacting an intermediate of formula 4 with an appropriate reducing agent, for example NaBH4 in an appropriate solvent(s) such as methanol (MeOH) and tetrahydrofuran (THF). Reaction of the resulting intermediate with a suitable alkylating agent, such as R2W, wherein R2 is as defined above and W is an appropriate leaving group, such as halo. Said reaction requires the presence of a base, such as aqueous KOH. The scheme 3 method is particularly useful for compounds of formula 1
wherein R1 is different than R2. 
Appropriate substituents, such as free nitrogen atoms, of the compounds of formula 1 may be protected with an optional protective group. These protective groups can be removed after the reactions or transformations described above. For example, in the imidazole moiety of a R1 group, a free nitrogen atom may be protected with a triphenylmethyl (trityl) group, which may be removed by stirring in the presence of trifluoroacetic acid (TFA) and triethylsilane.
The substituents of the compounds of formula 1 may be converted to other substituents falling within the scope of formula 1 via reactions or functional group transformations familiar to those skilled in the art. A number of such transformations are already described above. For example, an aldehyde functional group may be transformed into a cyano functional group by methods known to those skilled in the art, such as reacting the compound of formula 1 with hydroxylamine hydrochloride in an appropriate solvent such as a mixture of dichloromethane (DCM) and an alcohol, such as ethanol, isolating a product, and then stirring the product in the presence of triethylamine and p-toluenesulphonyl chloride.
Other examples are hydrolysis of carboxylic esters to the corresponding carboxylic acid or alcohol; hydrolysis of amides to the corresponding carboxylic acids or amines; hydrolysis of nitriles to the corresponding amides; amino groups on imidazole or phenyl moieties may be replaced by hydrogen by diazotation reactions familiar to those skilled in the art, and subsequent replacement of the diazo-group by hydrogen; alcohols may be converted into esters and ethers; primary amines may be converted into secondary or tertiary amines; double bonds may be hydrogenated to the corresponding single bond.
With reference to Scheme 4 below, the compound of formula 5, wherein W is an appropriate leaving group, such as halo, can be reacted to add an R4 group, wherein R4 is an aryl group or a heterocyclic aryl group and using coupling reagents that are known to those of skill in the art, such as palladium catalysis (with a palladium reagent, such as tetrakis(triphenylphosphine)palladium(0)) in the presence of a coupling partner of formula ZR4, wherein Z is an appropriate coupling group, such as xe2x80x94B(OH)2. 
With reference to Scheme 5 below, compounds of formula 3, wherein the R1 and R2 substituents are the same and are designated by R, may be prepared as described above in Scheme 1 for the preparation of a compound of formula 1, by alkylating a compound of formula 5, according to methods familiar to those skilled in the art, such as by stirring the intermediate of formula 6 in the presence of a base and an alkylating agent in an appropriate solvent. For example, an appropriate base is potassium hexamethyldisilazide; an appropriate solvent is, for example, tetrahydofuran; an appropriate alkylating agent is Rxe2x80x94Z, wherein Z is a leaving group, such as chloro. 
With reference to Scheme 6 below, the compound of formula 7 may be reacted in the presence of an acid, such as trifluoroacetic acid (TFA) to form a compound of formula 5a, which are intermediates of compounds of formula 5, wherein n is 0. The compound of formula 7 may be prepared by cyclizing a compound of formula 6 in the presence of an acylating agent, such a triphosgene, in an appropriate solvent, such as DCM, and in the presence of a base, such as diisopropylethyl amine, and optionally in the presence of a hindered base, such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). The resulting compound of formula 7, wherein R3 is hydrogen, may be converted to compounds of formula 7, wherein R3 is other than hydrogen as defined above, using procedures familiar to those skilled in the art. 
With reference to Scheme 7 below, compounds of formula 3a, which are intermediates of formula 3, wherein n is 0, may be prepared by reducing a compound of formula 11, according to methods familiar to those skilled in the art, such as reacting a compound of formula 11 in the presence of an appropriate reducing agent, such as a hydride reducing agent (such as sodium borohydride), in an appropriate solvent, such as a mixture of tetrahydrofuran and an alcohol, such as methanol (converting the alkenyl-R substituent into either a R1 or a R2 group), and then reacting the reduction product with a reagent of formula RZ, wherein Z is an appropriate leaving group and R is either R1 or R2, under appropriate conditions. Compounds of formula 11 may be prepared by reacting a compound of formula 10 with a reagent of formula Hxe2x80x94R1 or Hxe2x80x94R2, in the presence of suitable reagent(s), such as acetic acid and acetic anhydride. Compounds of formula 10 may be prepared by reacting a compound of formula 9 under conditions that are familiar to those skilled in the art, such as in the presence of sulfuric acid. Compounds of formula 9, wherein R3 is hydrogen, may be prepared by reacting a compound of formula 8 with ethylene glycol in the presence of an acid, such as p-toluenesulfonic acid, and an appropriate solvent, such as benzene, at reflux temperatures with the removal of water. The resulting compound of formula 9, wherein R3 is hydrogen, may be converted to compounds of formula 9, wherein R3 is other than hydrogen as defined above, using procedures familiar to those skilled in the art.
Compounds of formula 9 may be protected with a protecting group other than dioxolane group used in the methods of Scheme 5. These protecting groups are familiar to those skilled in the art. When the compounds of formula 9 are protected with a protecting group other than dioxolane, those compounds may be converted to compounds of formula 10 according to methods familiar to those skilled in the art. 
With reference to Scheme 8 below, the compounds of formula 4 which are those wherein n is 0, wherein the alkenyl-R substituent is R1 or R2, may be prepared by reacting a compound of formula 13 with a compound of formula RC(O)H in a suitable solvent, such as methanol, in the presence of a base such as pyrrolidine, at a temperature ranging from room temperature to reflux, preferably at about 60 EC. Compounds of formula 13 may be prepared from a compound of formula 12 using methods similar to those described in Scheme 4 above to prepare compounds of formula 2. 
With reference to Scheme 9 below, compounds of formula 7 wherein each W is an appropriate leaving group, such as halo, and R is an appropriate carboxylic acid protecting group, such as tert-butyl, may be prepared by cyclizing a compound of formula 16 in the presence of an appropriate reagent, such as triphosgene, in an inert solvent, such as DCM, and in the presence of a base, such as diisopropylethyl amine, and optionally in the presence of a hindered base, such as DBU. The resulting compound of formula 7, wherein R3 is hydrogen, may be converted to compounds of formula 7, wherein R3 is other than hydrogen as defined above, using procedures familiar to those skilled in the art. Compounds of formula 16 may be prepared from compounds of formula 15 using catalytic hydrogenation conditions, such as by using platinum on carbon in a reaction-inert solvent such as ethanol in the presence of H2. Compounds of formula 15 may be prepared by reacting a compound of formula 14 with a malonate derivative, such as di-tert-butyl malonate, in the presence of a base, such a sodium hydride, in an appropriate solvent, such as dimethylsulfoxide, at a temperature ranging from room temperature to 160 EC, preferably at about 100 EC. 
With reference to Scheme 10 below, an alternative method of preparing compounds of formula 5a, which are compounds of formula 5, wherein n is 0, and the R3 group is bound to the oxindole nitrogen via an alkyl group, begins with a compound of formula 6 which may be reacted with a compound of formula 17, in the presence of a reducing agent, such as sodium triacetoxyborohydride, in an appropriate solvent, such as acetic acid. 
With reference to Scheme 11 below, an alternative method of preparing compounds of formula 5a, begins with a compound of formula 18 (wherein W is a leaving group, such as halo) which may be reacted in the presence of hydroxylamine, chloral hydrate, sodium sulfate, an acid, such as HCl, and in an aqueous solution to give a compound of formula 19. Compounds of formula 20 may be formed by reacting a compound of formula 19 in the presence of an acid, such as sulfuric acid (see, for example, Synthesis, p. 993 (1993); J. Med. Chem., 29, p. 648 (1986)). Compounds of formula 20 may be converted to compounds of formula 21, wherein R3 is other than hydrogen as defined above, using procedures familiar to those skilled in the art. Compounds of formula 21 may be converted to compounds of formula 5a under reductive conditions that are familiar to those skilled in the art, such as in the presence of hydrazine hydrate at an elevated temperature, preferably from about 50 to 120xc2x0 C. (see, for example, Syn. Comm., 24, p. 2835 (1994). 
With respect to Scheme 12 below, compounds of formula 5b, which are compounds of formula 5 wherein n is 1, may be prepared starting from compounds having formula 22. Compounds having formula 23 may be prepared from compounds having formula 22 by methods familiar to those skilled in the art, such as reacting the acid compound in the presence of an appropriate reagent, such as thionyl chloride. Reaction of the compound of formula 23 in the presence of a reagent, such as lead thiocyanate leads to a compound having formula 24, which may be cyclized in the presence of a Lewis acid, such as AlCl3 in an appropriate solvent, such as carbon disulfide (CS2) to form a compound of formula 25. Conversion of the compound of formula 25 into the compound having formula 26 may be carried out using methods familiar to skilled practitioners, such as stirring the compound of formula 25 in a solution of aqueous base (see, J. Org. Chem., 29 p. 2261 (1964)). Compounds of formula 27 may be prepared by stirring compounds of formula 26 in the presence of a dehydrating agent, such as dicyclohexylcarbodiimide (DCC), in an appropriate solvent, such as acetonitrile. Reaction of compounds of formula 27 in the presence of an amine of formula R3xe2x80x94NH2, wherein R3 is as defined above, in an appropriate solvent(s) such as xylene/dioxane, at an elevated temperature, preferably from about 70 to 150xc2x0 C. provides compounds of formula 5b. 
The compounds of formula 1 and some of the intermediates described above may have one or more stereogenic centers in their structure. Such stereogenic centers may be present in a R or a S configuration. Oxime moieties, such as where R5 is xe2x80x94CHxe2x95x90NOR7, may exist in E or Z configurations.
The compounds of formula 1 as prepared in the above processes may be racemic mixtures of enantiomers which can be separated from one another following resolution procedures familiar to those skilled in the art. The racemic compounds of formula 1 may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali. An alternative manner of separating the enantiomeric forms of the compounds of formula 1 involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound will be synthesized by stereospecfic methods of preparation. These methods will advantageously employ enantiomerically pure starting materials.
The compounds of formula 1 that are basic in nature are capable of forming a wide variety of different salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often desirable in practice to initially isolate the compound of formula 1 from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert the latter back to the free base compound by treatment with an alkaline reagent and subsequently convert the latter free base to a pharmaceutically acceptable acid addition salt. The acid addition salts of the base compounds of this invention are readily prepared by treating the base compound with a substantially equivalent amount of the chosen mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent, such as methanol or ethanol. Upon evaporation of the solvent, the desired solid salt is readily obtained. The desired acid addition salt can also be precipitated from a solution of the free base in an organic solvent by adding to the solution an appropriate mineral or organic acid. Cationic salts of the compounds of formula 1 are similarly prepared except through reaction of a carboxy group with an appropriate cationic salt reagent, such as sodium, potassium, calcium, magnesium, ammonium, N,Nxe2x80x2-dibenzylethylenediamine, N-methylglucamine (meglumine), ethanolamine, tromethamine, or diethanolamine.
The compounds of formula 1 and their pharmaceutically acceptable salts, prodrugs, and solvates (hereinafter referred to, collectively, as xe2x80x9cthe therapeutic compoundsxe2x80x9d) can be administered orally, transdermally (e.g., through the use of a patch), parenterally or topically. Oral administration is preferred. In general, compounds of the formula 1 and their pharmaceutically acceptable salts, prodrugs, and solvates are most desirably administered in dosages ranging from about 1.0 mg up to about 500 mg per day, preferably from about 1 to about 100 mg per day in single or divided (i.e., multiple) doses. The therapeutic compounds will ordinarily be administered in daily dosages ranging from about 0.01 to about 10 mg per kg body weight per day, in single or divided doses. Variations may occur depending on the weight and condition of the person being treated and the particular route of administration chosen. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day.
The therapeutic compounds may be administered alone or in combination with pharmaceutically acceptable carriers or diluents by either of the two routes previously indicated, and such administration may be carried out in single or multiple doses. More particularly, the novel therapeutic compounds of this invention can be administered in a wide variety of different dosage forms, i.e., they may be combined with various pharmaceutically acceptable inert carriers in the form of tablets, capsules, lozenges, troches, hard candies, powders, sprays, creams, salves, suppositories, jellies, gels, pastes, lotions, ointments, elixirs, syrups, and the like. Such carriers include solid diluents or fillers, sterile aqueous media and various non-toxic organic solvents, etc. Moreover, oral pharmaceutical compositions can be suitably sweetened and/or flavored.
For oral administration, tablets containing various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine may be employed along with various disintegrants such as starch (and preferably corn, potato or tapioca starch), alginic acid and certain complex silicates, together with granulation binders like polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for tabletting purposes. Solid compositions of a similar type may also be employed as fillers in gelatin capsules; preferred materials in this connection also include lactose or milk sugar as well as high molecular weight polyethylene glycols. When aqueous suspensions and/or elixirs are desired for oral administration, the active ingredient may be combined with various sweetening or flavoring agents, coloring matter or dyes, and, if so desired, emulsifying and/or suspending agents as well, together with such diluents as water, ethanol, propylene glycol, glycerin and various like combinations thereof.
For parenteral administration, solutions of a therapeutic compound in either sesame or peanut oil or in aqueous propylene glycol may be employed. The aqueous solutions should be suitably buffered if necessary and the liquid diluent first rendered isotonic. These aqueous solutions are suitable for intravenous injection purposes. The oily solutions are suitable for intra-articular, intramuscular and subcutaneous injection purposes. The preparation of all these solutions under sterile conditions is readily accomplished by standard pharmaceutical techniques well-known to those skilled in the art.
Additionally, it is also possible to administer the therapeutic compounds topically and this may preferably be done by way of creams, jellies, gels, pastes, ointments and the like, in accordance with standard pharmaceutical practice.
The therapeutic compounds may also be administered to a mammal other than a human. The dosage to be administered to a mammal will depend on the animal species and the disease or disorder being treated. The therapeutic compounds may be administered to animals in the form of a capsule, bolus, tablet or liquid drench. The therapeutic compounds may also be administered to animals by injection or as an implant. Such formulations are prepared in a conventional manner in accordance with standard veterinary practice. As an alternative the therapeutic compounds may be administered with the animal feedstuff and for this purpose a concentrated feed additive or premix may be prepared for mixing with the normal animal feed.
The compounds of formula 1 exhibit activity as Ras farnesylation inhibitors and are useful in the treatment of cancer and the inhibition of abnormal cell growth in mammals, including humans. The activity of the compounds of formula 1 as Ras famesylation inhibitors may be determined by their ability, relative to a control, to inhibit Ras farnesyl transferase in vitro. This procedure is described below.
A crude preparation of human farnesyl transferase (FTase) comprising the cytosolic fraction of homogenized brain tissue is used for screening compounds in a 96-well assay format. The cytosolic fraction is prepared by homogenizing approx. 40 grams fresh tissue in 100 ml of sucrose/MgCl2/EDTA buffer (using a Dounce homogenizer; 10-15 strokes), centrifuging the homogenates at 1000 g for 10 minutes at 4xc2x0 C., re-centrifuging the supernatant at 17,000 g for 15 minutes at 4xc2x0 C., and then collecting the resulting supernatant. This supernatant is diluted to contain a final concentration of 50 mM Tris HCl (pH 7.5), 5 mN DTT, 0.2 M KCl, 20 mM ZnCl2, 1 mM PMSF and re-centrifuged at 178,000 grams for 90 minutes at 4 G. The supernatant, termed xe2x80x9ccrude FTasexe2x80x9d was assayed for protein concentration, aliquoted, and stored at xe2x88x9270xc2x0 C.
The assay used to measure in vitro inhibition of human FTase is a modification of the method described by Amersham LifeScience for using their Farnesyl transferase (3H) Scintillation Proximity Assay (SPA) kit (TRKQ 7010). FTase enzyme activity is determined in a volume of 100 ml containing 50 mM N-(2-hydroxy ethyl) piperazine-N-(2-ethane sulfonic acid) (HEPES), pH 7.5, 30 mM MgCl2, 20 uM KCl, 5 mM Na2HPO4, 5 mM dithiothreitol (DTT), 0.01% Triton X-100, 5% dimethyl sulfoxide (DMSO), 20 mg of crude FTase, 0.12 mM [3H]-farnesyl pyrophosphate ([3H]-FPP; 36000 dpm/pmole, Amersham LifeScience), and 0.2 mM of biotinylated Ras peptide KTKCVIS (Bt-KTKCVIS) that is N-terminally biotinylated at its alpha amino group and was synthesized and purified by HPLC in house. The reaction is initiated by addition of the enzyme and terminated by addition of EDTA (supplied as the STOP reagent in kit TRKQ 7010) following a 45 minute incubation at 37xc2x0 C. Prenylated and unprenylated Bt-KTKCVIS is captured by adding 10 ml of steptavidin-coated SPA beads (TRKQ 7010) per well and incubating the reaction mixture for 30 minutes at room temperature. The amount of radioactivity bound to the SPA beads is determined using a MicroBeta 1450 plate counter. Under these assay conditions, the enzyme activity is linear with respect to the concentrations of the prenyl group acceptor, Bt-KTKCVIS, and crude FTase, but saturating with respect to the prenyl donor, FPP. The assay reaction time is also in the linear range.
The test compounds are routinely dissolved in 100% dimethyl sulfoxide (DMSO). Inhibition of farnesyl transferase activity is determined by calculating percent incorporation of tritiated-farnesyl in the presence of the test compound vs. its incorporation in control wells (absence of inhibitor). IC50 values, that is, the concentration required to produce half maximal farnesylation of Bt-KTKCVIS, is determined from the dose-responses obtained.
The following Examples further illustrate the invention. In the following Examples, xe2x80x9cEtxe2x80x9d refers to ethyl, xe2x80x9cMexe2x80x9d refers to methyl, and xe2x80x9cAcxe2x80x9d refers to acetyl.